SD大鼠骨髓间充质干细胞的生物学及电生理特性.docVIP

  SD 大鼠骨髓间充质干细胞的生物学及电生 理特性# 朱天琦1，孙敏娴2，张雪琴1，魏明发1** 5 10 15 20 25 30 35 40 （1. 武汉华中科技大学附属同济医院小儿外科，武汉 430000； 2. 武汉市中心医院内分泌科，武汉 430000） 摘要：目的：建立体外培养扩增、检测鉴定 SD 大鼠骨髓间充质干细胞(BMSCs)的方法，观 察其生物学及电生理特性；方法：采取全骨髓培养法分离和纯化大鼠 BMSCs，用显微镜观 察细胞形体，用流式细胞仪检测细胞纯度，观察 BMSCs 在诱导条件下向成骨细胞分化的能 力，采用全细胞膜片钳技术记录 BMSCs 细胞膜电流；结果：全骨髓培养法培养的 BMSCs 具有良好的贴壁性，流式细胞仪检测： CD90：95.63%，CD45：0.64%；第 4 代细胞经诱导 剂诱导可向成骨细胞分化；膜片钳记录到 3 种外向钾电流存在；结论：用全贴壁筛选法培养 出高纯度的 BMSCs，增殖稳定，在条件诱导下具有多分化的潜能。 关键词：SD 大鼠；骨髓间充质干细胞；细胞培养；膜片钳 中图分类号：R-332 Biological characteristics and electrophysiological property of bone marrow mesenchymal stern cells from SD rat in vitro Zhu Tianqi1, Sun Minxian2, Zhang Xueqin1, Wei Mingfa1 (1. pediatric surgery,Tongji hospital,Huazhong university of science and technology, WuHan 430000; 2. Department of Endocrinology,central hospital of wuhan city, WuHan 430000) Abstract: Object: To explore the approaches to isolate, culture and identify the bone marrow mesenchymal stem cells (BMSCs) of SD rat in vitro. Methods: The BMSCs were enriched and expanded by whole bone marrow culture. The morphology of the BMSCs was examined and analyzed by light microscopy and the purity and phemotype were analyzed by flow cytometry. The ability of osteogenic differentiation of the BMSCs under different induction conditions was assessed. The whole cell patch clamp technique was used to record the ionic currents in BMSC．Results: The BMSCs expanded by whole bone marrow culture were adherent cells with a similar spindle-shaped morphologh during different passages. The expressions of CD90 and CD45 were 95.63%、 0.64% respectively. The BMSCs could be induced to differentiate into osteoblasts. The voltage-clamp experiment showed that there were 3 types of outward currents existing in rat BMSCs．Conclusion: Rat BMSCs can be isolated and expanded by whole bone marrow culture, which is an efficient and simple method with high purity and good stability. Keywords: SD Rat; Bone marrow mesenchymal stem cell; Cell culture; Patch clamp 0 引言 骨髓